HIV PROTEASE INHIBITORS

Abstract: Compounds of formula (I) are HIV protease inhibitors. These compounds are useful in the prevention or treatment of infection by HIV and in the treatment of AIDS, either as compounds, pharmaceutically acceptable salts, pharmaceutical composition ingredients, whether or not in combination with other antivirals, immunomodulators, antibiotics or vaccines. Methods of treating AIDS and methods of preventing or treating infection by HIV are also described.

Description: TITLE OF THE INVENTION HIV PROTEASE INHIBITORS This application is a continuation-in-part of Merck 19139IA, filed December 15, 1993. This application is related to Merck 18996, U.S.S.N. 08/059,038, filed May 7, 1993, which is a continuation-in-part of Merck 18597IB, which is a continuation-in-part of Merck 18597IA, which is a continuation-in-part of U.S. Serial No. 07/789,508, filed November 8, 1991. This application is related to the following cases: U.S. Serial No. 595,913, filed October 11, 1990 (Merck Case 18236); U.S. Serial No. 746,460, filed August 16, 1991 (Merck case 18466); Merck case 18583, filed October 23, 1991; and Merck case 18416. The present invention is concerned with compounds which inhibit the protease encoded by human immunodeficiency virus (HIV), or pharmaceutically acceptable salts thereof, and are useful in the prevention of infection by HTV, the treatment of infection by HIV and the treatment of the resulting acquired immune deficiency syndrome (AIDS). It also relates to pharmaceutical compositions containing the compounds and to a method of use of the present compounds and other agents for the treatment of AIDS and viral infection by HTV. BACKGROUND OF THE INVENTION A retrovirus designated human immunodeficiency virus (HIV) is the etiological agent of the complex disease that includes progressive destruction of the immune system (acquired immune deficiency syndrome; AIDS) and degeneration of the central and peripheral nervous system. This virus was previously known as LAV, HTLV-III, or ARV. A common feature of retrovirus replication is the extensive post-translational processing of precursor polyproteins by a virally encoded protease to generate mature viral proteins required for virus assembly and function. Inhibition of this processing prevents the production of normally infectious virus. For example, Kohl, N.E. et al., Proc. Nat'l Acad. Sci.. 85. 4686 (1988) demonstrated that genetic inactivation of the HIV encoded protease resulted in the production of immature, non-infectious virus particles. These results indicate that inhibition of the HTV protease represents a viable method for the treatment of AIDS and the prevention or treatment of infection by HIV. The nucleotide sequence of HIV shows the presence of a gol gene in one open reading frame [Ratner, L. et al., Nature. 313. 277(1985)]. Amino acid sequence homology provides evidence that the pol sequence encodes reverse transcriptase, an endonuclease and an HIV protease [Toh, H. et aL, EMBO J.. 4, 1267 (1985); Power. M.D. et al, Science. 231. 1567 (1986); Pearl, L.H. et al.. Nature. 329. 351 (1987)]. Applicants demonstrate that compounds of this invention are inhibitors of HTV protease. BRIEF DESCRIPTION OF THE INVENTION Compounds of Formula I, as herein defined, are useful in the inhibition of HIV protease, the prevention of infection by HIV, the treatment of infection by HIV and in the treatment of AIDS, either as compounds, pharmaceutically acceptable salts, pharmaceutical composition ingredients, whether or not in combination with other antivirals, immunomodulators, antibiotics or vaccines. Methods of treating AIDS, methods of preventing infection by HIV, and methods of treating infection by HIV are also disclosed. Some abbreviations that may appear in this application are as follows. ABBREVIATIONS Designation Protecting Group BOC (Boc) t-butyloxycarbonyl CBZ (Cbz) benzyloxycarbonyl(carbobenzoxy) TBS (TBDMS) t-butyl-dimethylsilyl Activating Group HBT(HOBTorHOBt) 1 -hydroxybenzotriazole hydrate DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS This invention is concerned with compounds of Formula I, combinations thereof, or pharmaceutically acceptable salts thereof, in the inhibition of HIV protease, the prevention or treatment of infection by HIV and in the treatment of the resulting acquired immune deficiency syndrome (AIDS). Compounds of Formula I are defined as follows: or a pharmaceutically acceptable salt thereof, wherein: -O is a stable 8- to 10-membered bicyclic heterocycle, any ring of which may be saturated or unsaturated, and said heterocycle consists of carbon atoms and 1-3 heteroatoms selected from the group consisting of N, S or O, said heterocycle being unsubstituted or substituted with OH, halo, Cl-4 alkyl, oxo; with the proviso that neither One embodiment of the present invention is compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein: -O is a stable 8- to 10-membered bicyclic heterocycle, any ring of which may be saturated or unsaturated, and said heterocycle consists of carbon atoms and 2 heteroatoms selected from the group consisting of N or O, wherein the heteroatoms are in different rings. A second embodiment are compounds of Formula I wherein: -o is restricted to and X is O or S, or a pharmaceutically acceptable salt thereof. A third embodiment are compounds of Formula I wherein: is restricted to or pharmaceutically acceptable salts thereof Another embodiment of the present invention is compound which is N-(2(R)-hydroxy-l(S)-indanyl)-2(R)-phenylmethyl-4(S)- hydroxy-5-(l-(4-(3-furo[2,3-b]pyridylmethyl)-2(S)-N'-(t-butylcarbox- amido)-piperazinyl))pentaneamide, or pharmaceutically acceptable salt thereof. The compounds of the present invention, have chiral centers and occur as racemates, racemic mixtures and as individual diastereomers, or enantiomers with all isomeric forms being included in the present invention. A racemic mixture encompasses mixtures of stereoisomers of 50:50 and other ratios. When any variable (e.g., -α ^-^ ) occurs more than one time in any constituent or in Formula I, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. As used herein except where noted, "alkyl" is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms (Me is methyl, Et is ethyl, Pr is propyl, Bu is butyl); "Halo", as used herein, means fluoro, chloro, bromo and iodo. The pharmaceutically-acceptable salts of the compounds of Formula I (in the form of water- or oil-soluble or dispersible products) include the conventional non-toxic salts or the quaternary ammonium salts which are formed, e.g., from inorganic or organic acids or bases. Examples of such acid addition salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dihydrochloride, diphosphate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glutamate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy- ethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalene- sulfonate, nicotinate, nitrate, oxalate, pamoate, pectinate, persulfate, 3- phenylpropionate, phosphate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups may be quatemized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl; and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides and others. Other pharmaceutically acceptable salts include the sulfate salt ethanolate and sulfate salts. Schemes I-H for preparing the novel compounds of this invention are presented below. The examples specifically illustrate the application of the following schemes to specific compounds. Amide couplings used to form the compounds of this invention are typically performed by the carbodiimide method with reagents such as dicyclohexylcarbodiimide, or l-ethyl-3-(3-dimethyl- aminopropyl) carbodiimide. Other methods of forming the amide or peptide bond include, but are not limited to the synthetic routes via an acid chloride, azide, mixed anhydride or activated ester. Typically, solution phase amide coupling are performed, but solid-phase synthesis by classical Merrifield techniques may be employed instead. The addition and removal of one or more protecting groups is also typical practice. Additional related information on synthetic background is contained in EPO 0337714 and EPO 0541168. One method for producing Formula I compounds is provided by Scheme I. Dihydro-5(S)-(tert-butyldimethylsilyloxy- methyl)-3(2H)-furanone (Compound 1 below) is prepared by standard methods known in the art from commercially available dihydro-5(S)- (hydroxymethyl)-2(3H)-furanone. After alkylation of Compound 1 to form Compound 2, the protecting group of lactone 2 is removed with aqueous HF to afford Compound 3. The alcohol group of 3 is activated by conversion into a leaving group such as mesylate, tosylate or triflate by treating the alcohol with a sulfonyl chloride or (preferably) sulfonic anhydride, such as trifluoromethanesulfonic anhydride, in the presence of a hindered amine base such as triethylamine, diethyl isopropylamine or 2,6 lutidine, to afford a compound such as Compound 4. The leaving group of Compound 4 is displaced by an amine 5, such as 4-(l,l- dimethyl ethoxycarbonylamino)-piperazine-2(S)-carboxamide in a solvent such as DMF or xylene to produce a compound such as 6. A trifluoromethanesulfonyloxy group can be displaced by an amine at room temperature in a solvent such as isopropanol or methylene chloride by treatment with N,N-diisopropyl-ethylamine. Compound 6 is hydrolyzed with aqueous lithium or sodium hydroxide and the resultant hydroxy acid 7 is converted into a protected hydroxy acid 8. The hydroxyl group is conveniently protected with a standard silyl protecting group such as t-butyldimethyl silyl or t- butyldiphenyl silyl. The protected hydroxy-acid 8 is then coupled to the desired Rl2 amine to produce Compound 9, and the silyl protecting group is removed with fluoride ion to arrive at Compound 10. SCHEME I / ^ OH R3 T SiCI 2. MeOH Boc ^ N N C02 H N^ NH ' *CONH-|- SCHEME I (CONT'D a 10 10 20 25 30 SCHEME π 15 OH K 2 C0 3 25 30 SCHEME H (CONT'D) One preferred method is the synthesis of the epoxide 12 by reaction of H in the presence of strong base. The strong base must be a metal-containing base, in an inert anhydrous organic solvent, such as, e.g., cyclic or acyclic hydrocarbons including hexane, pentane, cyclohexane, etc. Suitable strong bases include: LiN[(CH3)3Si]2, KN[(CH3)3Si]2, NaN[(CH3)3Si]2, n-butyllithium (n-BuLi), s-BuLi, t- BuLi, potassium tert-butoxide, lithium diisopropyl-amide (LDA), lithium isopropylcyclohexylamide, lithium pyrrolidide, lithium tetramethylpiperidide, phenyllithium, isopropylmagnesium chloride, isobutylmagensmm chloride, and other similar strong base known in the art. Preferred strong bases are n-BuLi, s-BuLi, LiN[(CH3)3Si]2 and LDA, with n-BuLi and LiN[(CH3)3Si]2 being most preferred. Preferably, about 1 to 2 molar equivalents of strong base are used per 1 molar equivalent of 1_L Compound 13 is made by reacting Compound 12 with N-t- butyl-4-( 1 , 1 -dimethylethoxycarbonylamino)piperazine-2(S)- carboxamide (£). Preferably from about 1 to 3 molar equivalents of amine are used per molar equivalent of epoxide 12, with a ratio of about 1.05:1 molar equivalents of V:IV being more preferred. This reaction can be run in any suitable solvent, such as, e.g., one chosen from hydrocarbons, such as toluene, ethers such as di¬ ethyl ether, alcohols such as methanol, ethanol or isopropanol, nitriles such as acetonitriie, and esters such as ethyl acetate or combinations thereof, with alcohols being preferred and isopropanol being most preferred. The temperature of the reaction can be maintained in a range from ambient to the reflux temperature of the solvent used, but is preferably run at an elevated temperature, e.g., in the range of 80°C to 90°C, and most preferably from about 83°C to 85°C. Activated glycidols can be prepared by methods known in the art, such as described in, e.g., J. Klunder, et al-. J. Org. Chem.. 1989, 54 * 1295-1304 and references cited therein. Amide compounds such as J , can be made according to standard procedures known to those skilled in the art, such as, e.g., the procedure described in Example 10, using the appropriate starting materials. Protecting groups such as nitrogen protecting groups may be used where appropriate in the practice of this invention. For example, the 4 position nitrogen of 2-t-butylcarboxamide piperazine may be protected with a group such as BOC, CBZ, benzyl, 4-methoxy- benzyl, 2,4-dimethoxybenzyl, trifluoroacetamide, trialkylsilyl, or other groups known in the art. A compound of Formula 15 wherein P is a nitrogen protecting group such as -BOC or -CBZ, is also prepared according to the method described in Scheme I, preferably employing the 5-trif uoromethanesulfonyloxymethyl analog of lactone 4 therein. Compounds of Formula 16 can be obtained by a variety of routes from Compound 14 which is obtained after removal of the nitrogen protecting group in 15 using methods well known in the art, e.g., catalytic hydrogenation to remove a CBZ group, or treatment with trimethylsilyltriflate and 2,6 lutidine at about 0°C in a solvent such as CH2O2, or treatment with 6N HCI in isopropanol, to remove a BOC group. The 4-position piperazinyl nitrogen of Compound 14 can be alkylated with a compound of Formula Rl-X in a solvent such as DMF in the presence of Et3N at room temperature, wherein X is -Cl, Br or -I. Techniques for these procedures are well known to those skilled in the art. The compounds of this invention are also illustrated by the table of Example 3 below. The compounds of this invention are useful in the preparation and execution of screening assays for antiviral compounds. For example, the compounds of this invention are useful for isolating enzyme mutants, which are excellent screening tools for more powerful antiviral compounds. Furthermore, the compounds of this invention are useful in establishing or determining the binding site of other antivirals to HIV protease, e.g., by competitive inhibition. Thus the compounds of this invention are commercial products to be sold for these purposes. The compounds of the present invention are useful in the inhibition of HIV protease the prevention or treatment of infection by the human immunodeficiency virus (HIV) and the treatment of consequent pathological conditions such as AIDS. Treating AIDS or preventing or treating infection by HIV is defined as including, but not limited to, treating a wide range of states of HTV infection: AIDS, ARC (AIDS related complex), both symptomatic and asymptomatic, and actual or potential exposure to HIV. For example, the compounds of this invention are useful in treating infection by HIV after suspected past exposure to HIV by, e.g., blood transfusion, organ transplant, exchange of body fluids, bites, accidental needle stick, or exposure to patient blood during surgery. For these purposes, the compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles. Thus, in accordance with the present invention there is further provided a method of treating and a pharmaceutical composition for treating HIV infection and AIDS. The treatment involves administering to a patient in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof. These pharmaceutical compositions may be in the form of orally-administrable suspensions or tablets; nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories. When administered orally as a suspension, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners/flavoring agents known in the art. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants known in the art. When administered by nasal aerosol or inhalation, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art. The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally- acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid. When rectally administered in the form of suppositories, these compositions may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquidify and/or dissolve in the rectal cavity to release the drug. Dosage levels of the order of 0.02 to 5.0 or 10.0 grams- per-day are useful in the treatment or prevention of the above-indicated conditions, with oral doses two-to-five times higher. For example, infection by HTV is effectively treated by the administration of from 1.0 to 50 milligrams of the compound per kilogram of body weight from one to four times per day. In one preferred regimen, dosages of 100- 400 mg every six hours are administered orally to each patient. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy. The present invention is also directed to combinations of the HIV protease inhibitory compounds with one or more agents useful in the treatment of ADDS. For example, the compounds of this invention may be effectively administered, whether at periods of pre- exposure and/or post-exposure, in combination with effective amounts of the AIDS antivirals, immunomodulators, anti-infectives, or vaccines known to those of ordinary skill in the art. TABLE C ANTΓVTRALS Drug Name Manufacturer Indication AL-721 Ethigen ARC, PGL (Los Angeles, CA) HIV positive, AIDS Recombinant Human Triton Biosciences AIDS, Kaposi's Interferon Beta (Almeda, CA) sarcoma, ARC Acemannan Carrington Labs ARC (Irving, TX) (See also immunomodulators) Cytovene Syntex sight threatening CMV Ganciclovir (Palo Alto, CA) peripheral CMV retinitis d4T Bristol-Myers AIDS, ARC Didehydrodeoxy- (New York, NY) thymidine ddl Bristol-Myers AIDS, ARC Dideoxyinosine (New York, NY) EL10 Elan Corp, PLC HTV infection (Gainesville, GA) (See also immunomo dulators ) Drug Name Manufacturer Indication Trisodium Astra Pharm. CMV retinitis, HTV Phosphonoformate Products, Inc infection, other CMV (Westborough, MA) infections Dideoxycytidine; Hoffman-La Roche AIDS, ARC ddC (Nutley, NJ) Novapren Novaferon Labs, Inc. HIV inhibitor (Akron, OH) Diapren, Inc. (Roseville, MN, marketer) Peptide T Peninsula Labs AIDS Octapeptide (Belmont, CA) Sequence Zidovudine; AZT Burroughs Wellcome AIDS, adv, ARC AIDS, adv, ARC (Rsch. Triangle Park, pediatric AIDS, NC) Kaposi's sarcoma, asymptomatic HTV infection, less severe HIV disease, neurological involvement, in combination with other therapies. Drug Name Manufacturer Indication Ansamycin LM 427 Adria Laboratories ARC (Dublin, OH) Erbamont (Stamford, CT) Dextran Sulfate Ueno Fine Chem. AIDS, ARC, HTV Ind. Ltd. positive asymptomatic (Osaka, Japan) Virazole Viratek/ICN asymptomatic HIV Ribavirin (Costa Mesa, CA) positive, LAS, ARC Alpha Interferon Burroughs Wellcome Kaposi's sarcoma, (Rsch. Triangle HIV in combination Park, NC) w/Retrovir Acyclovir Burroughs Wellcome AIDS, ARC, asymptomatic HTV positive, in combination with AZT. Antibody which Advanced Biotherapy AIDS, ARC neutralizes pH Concepts labile alpha aberrant (Rockville, MD) Interferon in an immuno-adsorption column (Irving, TX) anti-virals) (New Orleans, LA) sarcoma, ARC, PGL Imuthiol Diethyl Merieux Institute AIDS, ARC Dithio Carbamate (Miami, FL) Alpha-2 Schering Plough Kaposi's sarcoma Interferon (Madison, NJ) w/AZT:AIDS Methionine- TNI Pharmaceutical AIDS, ARC Enkephalin (Chicago, IL) MTP-PE Ciba-Geigy Corp. Kaposi's sarcoma Muramyl- (Summit, NJ) Tripeptide ANTI-INFECTIVES Drug Name Manufacturer Indication Spiramycin Rhone-Poulenc cryptosporidial Pharmaceuticals diarrhea (Princeton, NJ) Intraconazole- Janssen Pharm. histoplasmosis; R51211 (Piscataway, NJ) cryptococcal meningitis Trimetrexate Warner-Lambert PCP OTHER Drug Name Manufacturer Indication Recombinant Human Ortho Pharm. Corp. severe anemia Erythropoietin (Raritan, NJ) assoc. with AZT therapy Megestrol Acetate Bristol-Myers treatment of (New York, NY) anorexia assoc. w/AIDS Total Enteral Norwich Eaton diarrhea and Nutrition Pharmaceuticals malabsorption (Norwich, NY) related to AIDS It will be understood that the scope of combinations of the compounds of this invention with AIDS antivirals, immunomodulators, anti-infectives or vaccines is not limited to the list in the above Table, but includes in principle any combination with any pharmaceutical composition useful for the treatment of AIDS. Certain compounds of Table C are the following: Compound B is 6-chloro-4-(S)-cyclopropyl-3,4-dihydro-4-((2- pyridyl)ethynyl)quinazolin-2(lH)-one; Compound C is(-) 6-chloro-4(S)- trifluoromethyl-1 ,2-dihydro-4(H)-3,l-benzoxazin-2-one; nevirapine is 1 l-cyclopropyl-5,1 l-dihydro-4-methyl-6H-dipyrido[3,2-t :2',3'- e][l,4]diazepin-6-one. Compounds B and C are synthesized by the methods of EP 0,569,083, herein incorporated by reference for this purpose. Nevirapine is synthesized by Klunder, J. M. et ah, J. Med. Chem.. 35, 1887 (1992); Hargrave, K. D. et al, J. Med Chem.. 34, 2231 (1991); Cohen, K. A. et al, J. Biol. Chem.. 266. 14670 (1991), all three references herein incorporated by reference. Preferred combinations are simultaneous or alternating treatments of an inhibitor of HIV protease and a non-nucleoside inhibitor of HTV reverse transcriptase. An optional third component in the combination is a nucleoside inhibitor of HIV reverse transcriptase, such as AZT, ddC or ddl. A preferred inhibitor of HIV protease is Compound A. Preferred non-nucleoside inhibitors of HIV reverse transcriptase include Compound B, Compound C or nevirapine. These combinations may have synergistic effects on limiting the spread of HTV. Preferred combinations include the following (1) Compound A, with a preferred non-nucleoside inhibitor of HTV reverse transcriptase, and, optionally, AZT or ddl or ddC; (2) Compound A, and any of AZT or ddl or ddC. Assay for Inhibition of Microbial Expressed HIV Protease Inhibition studies of the reaction of the protease expressed in Eschericia coli with a peptide substrate [Val-Ser-Gln-Asn- (betanapthyl)Ala-Pro-Ile-Val, 0.5 mg/mL at the time the reaction is initiated] were in 50 mM Na acetate, pH 5.5, at 30°C for 1 hour. Various concentrations of inhibitor in 1.0 μl DMSO were added to 25 μl of the peptide solution in water. The reaction is initiated by the addition of 15 μl of 0.33 nM protease (0.11 ng) in a solution of 0.133 M Na acetate pH 5.5 and 0.1% bovine serum albumin. The reaction was quenched with 160 μl of 5% phosphoric acid. Products of the reaction were separated by HPLC (VYDAC wide pore 5 cm C- 18 reverse phase, acetonitriie gradient, 0.1% phosphoric acid). The extent of inhibition of the reaction was determined from the peak heights of the products. HPLC of the products, independently synthesized, proved quantitation standards and confirmation of the product composition. Compound A showed IC50 values of about 0.27 nM. CELL SPREAD ASSAY Inhibition of the spread of HIV in cell culture was measured according to Nunberg, J. H. et al., J. Virol.. 65. 4887 (1991). In this assay, MT-4 T-lymphoid cells were infected with HTV-1 (wild- type, unless otherwise indicated) by using a predetermined inoculum, and cultures were incubated for 24 hours. At this time, <1% of the cells were positive by indirect immunofluorescence. Cells were then extensively washed and distributed into 96-well culture dishes. Serial twofold dilutions of inhibitor were added to the wells, and cultures were continued for 3 additional days. At 4 days postinfection, 100% of the cells in control cultures were infected. HIV-1 p24 accumulation was directly correlated with virus spread. The cell culture inhibitory concentration was defined as the inhibitor concentration in nanomoles/liter which reduced the spread of infection by at least 95%, or CIC95. The CIC95 for Compound A is 25 nM. INHIBITION OF VIRUS SPREAD A. Preparation of HIV-infected MT-4 cell Suspension. MT cells were infected at Day 0 at a concentration of 250,000 per ml with a 1 :1000 dilution of HIV-1 strain nib stock (final 125 pg p24/ml; sufficient to yield <1% infected cells on day 1 and 25- 100% on day 4). Cells were infected and grown in the following medium: RPMI 1640 (Whittaker BioProducts), 10% inactivated fetal bovine serum, 4 mM glutamine (Gibco Labs) and 1:100 Penicillin- Streptomycin (Gibco Labs). The mixture was incubated overnight at 37 °C in 5% C02 atmosphere. B. Treatment with Inhibitors A matrix of nanomolar range concentrations of the pairwise combinations is prepared. At Day 1, aliquots of 125 μl of inhibitors are added to equal volumes of HTV -infected MT-4 cells (50,000 per well) in a 96-well microtiter cell culture plate. Incubation is continued for 3 days at 37°C in 5% C02 atmosphere. C. Measurement of Virus Spread Using a multichannel pipettor, the settled cells are resuspended and 125 μl harvested into a separate microtiter plate. The supernatant is assayed for HIV p24 antigen. The concentration of HIV p24 antigen is measured by an enzyme immunoassay, described as follows. Aliquots of p24 antigen to be measured are added to microwells coated with a monoclonal antibody specific for HIV core antigen. The microwells are washed at this point, and at other appropriate steps that follow. Biotinylated HlV-specific antibody is then added, followed by conjugated streptavidin- horseradish peroxidase. A color reaction occurs from the added hydrogen peroxide and tetramethylbenzidine substrate. Color intensity is proportional to the concentration of HIV p24 antigen. Calculation of Degree of Synergy or Enhanced Inhibition When there is synergy pairwise combinations of inhibitors are found to exhibit markedly enhanced inhibition of virus spread, in comparison to each inhibitor alone, or in comparison to merely additive inhibition of each inhibitor. The data is processed as follows: fractional inhibitory concentration ratios (FIC) are calculated according to Elion, et ah, J. Biol. Chem.. 208. 477 (1954). The minimum sum of FICS, which is the maximum synergy, is determined for various pairwise combinations. The smaller the number, the greater the synergy. EXAMPLE 1 Preparation of N-(2(R)-hydroxy-l(S)-indanyl)-2(R)-phenylmethyl- 4(S)-(hydroxy)-5-(l-(2(S)-N-(t-butyl-carboxamido)-piperazinyl)- pentaneamide. Compound 14 Step 1: Preparation of dihydro-5(S)-((t-butyldiphenylsilyl)- oxymethyl)-3(R)phenylmethyl-3(2H)-furanone A solution of lithium diisopropylamide (LDA) was generated by the addition 1.55 ml of n-BuLi (2.5 M in hexane) to 0.55 ml (3.9 mmol) of diisopropylamine in 10 ml of THF at -78°C. After 30 minutes a solution of dihydro-5-(S)-((t-butyldiphenylsilyl)-oxymethyl)- 3(2H)-furanone (1.38 g, 3.89 mmol) in 5 ml of THF was added. After an additional 30 minutes of stirring, benzyl bromide (0.68 g, 3.9 mmol) was added and stirring was continued for 3 hours after which time the reaction was quenched with the addition of a 10% aqueous citric acid solution. The solution was extracted with ethyl acetate (2 x 50 ml) which was backwashed with brine, dried, filtered and concentrated to afford an oil. The product was purified by chromatography (Siθ2, 20% EtOAc/Hexane) to afford the title compound. Step 2: Preparation of dihydro-5(S)-(hydroxy-methyl)-3(R)- phenylmethyl-3(2H)-furanone To 5.26 g of dihydro-5(S)-((t-butyldiphenylsilyl)oxy- methyl)-3(R)phenylmethyl-3(2H)-furanone in 40 ml of acetonitriie was added 1.34 ml of a 49% aqueous HF solution. After 18 hours at room temperature the reaction was concentrated to dryness and the residue was partitioned between water (50 ml) and ethyl acetate (50 ml). The organic layer was washed with brine, dried filtered and concentrated to afford the product as a tan solid (mp 69-72°C). Step 3: Preparation of dihydro-5(S)-((trifluoromemanesulfonyl)- oxymethyl)-3(R)-phenylmethyl-3(2H)-furanone To a solution of 18.4 g (89.2 mmol) of dihydro-5(S)- (hydroxymethyl)-3(R)-phenylmethyl-3(2H)-furanone in 350 mL of methylene chloride cooled to 0°C was added 13.51 mL 2,6-lutidine (115.98 mmol) followed by a dropwise addition of 16.51 mL of trifluoromethanesulfonic anhydride (98.1 mmol). After 1.5 hours at 0°C, the reaction was poured into a mixture of 300 mL ice/brine and stirred for 0.5 hours. The aqueous layer was then extracted with methylene chloride (3 x 150 mL), the organic layers were washed with 10% HCI (2 x 75 mL), saturated NaHCθ3 (100 mL), water (100 mL), dried over MgS04, filtered and concentrated to give a solid residue. Purification via flash column chromatography (120 x 150 mm column, gradient elution of hexanes :EtO Ac, 4:1 to 3:1) afforded the title product; mp 53-54°C. Step 4: Preparation of 4-( 1 , 1 -dimethylethoxy carbonylamino)- 1 - (phenylmethyicarbonylamino)-piperazine-2S-carboxylic acid The title compound was prepared following the procedure of Bigge, C.F.; Hays, S.J.; Novak, P.M.; Drummond, J.T.; Johnson, G.; Bobovski, T.P., Tetrahedron Lett.. 1989, 30, 5193; starting with 2(S)- piperazine-carboxylic acid, (see Felder, E.; Maffei, S.; Pietra, S.; Pitre, D., Helv. Chim. Acta. 1960, I V∑, 888. Step 5: Preparation of N-t-butyl-4-(l,l-dimethylethoxycarbonyl- amino)- 1 -(phenylmethylcarbonyl-amino)piperazine-2(S)- carboxamide To 9.90 g (27.16 mmol) of the product of Step 4 dissolved in 75 mL of DMF and cooled to 0°C was added 5.73 g (29.88 mmol) of EDC, 4.03 g (29.88 mmol) of HOBt, 3.14 mL (29.88 mmol) of t- butylamine, and finally 4.16 mL (29.88 mmol) of triethylamine. The reaction mixture was stirred for 18 hours and the reaction volume was concentrated by half. The mixture was then diluted with 600 mL of EtOAc and washed with 10% HCI (2 x 75 mL), saturated NaHCθ3 (1 x 75 mL), water (3 x 75 mL) and brine (1 x 50 mL), dried over MgS04 and concentrated to a solid. This solid was triturated with EtOAc: hexane (1:2) and filtered to provide the title product as a white solid; mp 134-135°C. Step 6: Preparation of N-t-butyl-4-( 1 , 1 -dimethylethoxycarbonyl- amino)piperazine-2(S)-carboxamide To 1.20 g (2.86 mmol) of N-t-butyl-4-(l,l-dimethyl- ethoxy-carbony lamino)- 1 -(phenylmethylcarbonylamino)piperazine- 2(S)-carboxamide and 1.1 g (0.086 mmol) of 10% Pd/C was added 15 mL of methanol. The vessel was charged with hydrogen and the reaction stirred for 2 hours, filtered through celite and washed with ethanol. The solvents were removed in vacuo to provide the title product as a foam. lH NMR (300 MHz, CDCI3) δ 6.65 (br, IH), 4.10 (m, IH), 3.81 (br, IH), 3.21 (dd, J=18 and 7 Hz, IH), 3.02-2.70 (m, 4H), 2.10-2.0 (br, IH), 1.50 (s, 9H), 1.41(s, 9H). Step 7: Preparation of dihydro-5(S)-(4-(l,l-dimethylethoxy- carbonylamino))-2(S)-N-(t-butylcarboxamido)- piperazinyl)methyl)-3(R)-phenylmethyl-3(2H)-mranone To a solution of 22.40 g (0.0662 mol) dihydro-5(S)- ((trifluoromethanesulfonyl)oxymethyl)-3(R)-phenylmethyl-3(2H)- furanone (prep in Step 3) and 18.0 g (0.063 mol) of N-t-butyl-4-(l,l- dimethylethoxycarbonylamino)piperazine-2(S)-carboxamide dissolved in 180 mL of isopropanol was added 11.53 mL (0.0662 mol) of NJSf- diisopropylethylamine. After 2.5 hours another 1.2 g of dihydro-5(S)- ((trifluoromethanesulfonyl)oxymethyl)-3(R)-phenylmethyl-3(2H)- furanone was added. The reaction was complete by thin layer chromatography (TLC) after 3.5 hours and was concentrated to a thick oil. Trituration with EtOAc:hexanes (1:2, 200 mL) provided a white solid which was filtered and discarded. The oil was purified by flash column chromatography (120 x 150 mm column, EtOAc:hexanes gradient elution 1:1, 2:1, 3:1 to all EtOAc) to afford the title compound. lH NMR (400 MHz, CDCI3) δ 7.34-7.17 (m, 5H), 6.31 (br s, IH), 4.38 (br m, IH), 3.96-3.92 (m, IH), 3.79 (br m, IH), 3.16 (dd, J=13.6 and 4.4 Hz, IH), 3.08-2.99 (m, 3H), 2.90-2.82 (m, IH), 2.80 (dd, J=13.5 and 8.9 Hz, IH), 2.78 (m, IH), 2.67-2.61 (m, IH), 2.58-2.49 (m, IH), 2.38-2.32 (m, IH), 2.32-2.04 (m, IH), 1.99-1.92 (m, IH), 1.45 (s, 9H), 1.29 (s, 9H). Step 8: Preparation of 2(R)-phenylmethyl-4(S)-(t-butyldimethyl- silyloxy)-5-(l -(4-(l ,1-di-methylethoxycarbonylamino)))- 2(S)-N-(t-butylcarboxamido)-piperazinyl))-pentaneamide To 25.50 g (52.50 mmol) of dihydro-5(S)-(4-(l,l- dimethyl-ethoxycarbonylamino))-2(S)-N-(t-butylcarboxamido)- piperazinyl)-methyl)-3(R)-phenylmethyl-3(2H)-furanone dissolved in 120 mL DME cooled to 0°C was added a solution of 60 mL of water and 1.512 g (63.01 mmol) of lithium hydroxide. After 0.5 hours the reaction was quenched with the addition of 10% HCI until pH 6 and the solution was concentrated jn vacuo. The residue was dissolved in 50 mL water and extracted with EtOAc (4 x 75 mL) and the organic layers were washed with water (1 x 20 mL), brine (1 x 20 mL). The aqueous was back extracted with EtOAc (2 x 75 mL) and the combined organic layers were dried over MgSθ4 and concentrated to provide a yellow solid. This crude product was dissolved in 100 mL of DMF and 17.87 g (0.262 mol) of imidazole was added, cooled to 0°C and then 31.50 g (0.21 mol) of t-butyldimethylsilyl chloride was added. This stirred 1 hour at 0°C and was then warmed to room temperature. After 20 hours the reaction was quenched with 10 mL methanol and concentrated to half the volume. 100 mL of pH 7 buffered water was added and the aqueous was extracted with EtOAc (4 x 100 mL), the combined organic layers were washed with 10% HCI (2 x 50 mL), water (3 x 75 mL), and brine (1 x 50 mL), dried over MgSθ4 and concentrated to obtain the title compound. This material was used directly in the next step. Step 9: Preparation of N-(2(R)-hy droxy- 1 (S)-indany 1)-2(R)- phenylmethyl-4(S)-(t-butyldimethylsilyloxy)-5-( 1 -(4-(l , 1 - dimethylethoxycarbonylammo)))-2(S)-N-(t-butylcarbox- amido)-piperazinyl))-pentaneamide To 27.0 g (0.0446 mol) of the crude material from Step 6 dissolved in 180 mL of DMF and cooled to 0°C was added 8.98 g (0.0468 mol) of EDC, 6.32 g (0.0468 mol) of HOBt, and 7.31 g (0.049 mol) aminohydroxy indane. Triethylamine (6.52 mL, 0.0468 mol) was added and the reaction stirred at 0°C for 2 hours, room temperature for 16 hours and was quenched by diluting with 500 mL of EtOAc. The organic layer was washed with 10% HCI (2 x 100 mL), saturated NaHCθ3 (1 x 100 mL), water (3 x 150 mL), brine (1 x 75 mL), dried over MgSθ4 and concentrated to yield the title compound as a white foam. lH NMR (400 MHz, CDCI3) δ 7.4-7.17 (m, 9H), 6.51 (br s, IH), 5.79 (br s, IH), 5.23 (m, IH), 4.23 (br s, IH), 4.06 (m, IH), 3.96-3.84 (m, 2H), 3.07-2.78 (m, 8H), 3.65 (dd, J=9.6 and 4.1 Hz, IH), 2.56-2.44 (m, 2H), 2.29 (dd, J=12.0 and 4.5 Hz, IH), 2.17-2.09 (m, IH), 1.79 (br s, IH), 1.44 (s, 9H), 1.35 (s, 9H), 1.10 (s, IH), 0.84 (s, 9H), 0.12 (s, 3H), 0.08 (s, 3H). Step 10: Preparation of N-^R^hydroxy-US^indanyl^W- phenylmethyM SMhydroxy^-O-^-QJ-dimethyl- ethoxycarbonylamino)))-2(S)-N-(t-but lcarboxamido)- piperazinyl))-pentaneamide To 32.20 g (0.0437 mol) of N-(2(R)-hydroxy-l-(S)- indanyl)-2(R)-phenylmethyl-4(S)-(t-butyldimethylsilyloxy)-5-(l-(4- (1,1 -dimethylethoxycarbonylamino)))-2(S)-N-(t-butylcarboxamido)- piperazinyl))-pentaneamide was added 437 mL (0.437 mol) of tetrabutylammonium fluoride (1.0 M solution in THF, Aldrich). The reaction stirred for 18 hours and was then concentrated to 200 mL and diluted with 700 mL of EtOAc. This was washed with water (2 x 100 mL), brine (1 x 50 mL) and the aqueous layers were back extracted with EtOAc (2 x 200 mL). The combined organic layers were dried over MgSθ4 and concentrated to an oil. Purification via flash column chromatography (120 x 150 mm column, gradient elution CH2CI2: CHCl3/saturated with NH3:methanol, increasing methanol from 1%, 1.5%, 2%) afforded the title compound as a white foam. lH NMR (400 MHz, CDCI3) δ 7.31-7.11 (m, 9H), 6.41 (br s, IH), 6.23 (d, J=8.6 Hz, IH), 5.25 (dd, J=8.6 and 4.7 Hz, IH), 4.21 (m, IH), 3.83- 3.82 (m, 2H), 3.78-3.61 (m, 2H), 3.22-3.19 (m, 2H), 3.03-2.78 (m, 8H), 2.62-2.58 (m, IH), 2.41-2.35 (m, 2H), 2.04-2.02 (m, IH), 1.57-1.50 (m, IH), 1.45 (s, 9H), 1.32 (s, 9H). Step 11 : Preparation of N-(2(R)-hydroxy-l(S)-indanyl)-2(R)- phenylmethyl-4(S)-(hydroxy)-5-(l-(2(S)-N-(t-butyl- carboxamido)-piperazinyl)-pentaneamide. Compound 14 To 21.15 g (0.034 mol) of N-(2(R)-hydroxy-l(S)-indanyl)- 2(R)-phenylmethyl-4(S)-(hydroxy)-5-(l-(4-(l,l-dimethylethoxy- carbonylamino)))-2(S)-N-(t-butylcarboxamido)-piperazinyl))- pentaneamide dissolved in 350 mL of methylene chloride and cooled to 0°C was added 22.43 mL (0.204 mol) 2,6-lutidine and then 32.85 mL (0.170 mol) of trimethylsilyltriflate over 5 minutes. After 0.5 hours the reaction was quenched with 10% HCI (80 mL) and this stirred 0.5 hours. To this was added 100 mL of saturated NaHCθ3 and then solid NaHCθ3 until pH 8. The aqueous layer was then extracted with EtOAc (4 x 100 mL) and the combined organic layers were washed with water (1 x 50 mL), brine (1 x 75 mL), dried over MgS04 and concentrated. The residue was purified via column chromatography (120 x 150 mm column, gradient elution CH2Cl2:CHCl3 saturated with NH3:MeOH, slowly increasing methanol 2%, 3%, 4%, 5%, 6%, to 10%). This provided the title product as a white foam. lH NMR (400 MHz, CDCI3) δ 7.53 (s, IH), 7.29-7.09 (m, 9H), 6.52 (d, J=8.3 Hz, IH), 5.24 (dd, J=8.2 and 4.9 Hz, IH), 4.23 (dd, J=4.7 and 4.03 Hz, IH), 4.25-4.00 (br s, IH), 3.83-3.81 (m, IH), 3.03-2.88 (m, 4H), 2.82-2.73 (m, 7H), 2.50-1.60 (br s, 2H), 2.45 (d, J=6.2 Hz, 2H), 2.32-2.29 (m, IH), 1.98 (m, IH), 1.51 (m, IH), 1.33 (s, 9H). EXAMPLE 2 Preparation of N-(2(R)-hydroxy-l(R)-indanyl)-2(R)-phenylmethyl- 4(R)-hydroxy-5-(l-(4-(3-furo[2,3-b]pyridylmethyl)-2(R)-N * -(t- butylcarboxamido)-piperazinyl))-pentaneamide Step 1: Preparation of furor2.3-blpyridine-2.5-dicarboxylic acid To solution of the known [Snyder, H.R., Ebetino, F.F., L Het. Chem.. 3, 202-205 (1966)] diethyl furo[2,3-b]pyridine-2,5- dicarboxylate (1.22 g, 4.923 mmol) in 10 mL of 95% ethanol was added a solution of potassium hydroxide (0.66 g, 11.81 mmol) dissolved in 10 mL of water. The reaction was warmed to 80°C for 3 hours, cooled to RT and filtered. The bispotassium salt was dissolved in water and acidified with 10% HCI to pH 2. The precipitate was filtered and dried under vacuum to afford 850 mg of a white solid. lH NMR (400 MHz, (CD3)2SO) δ 8.98 (d, J=2.2 Hz, IH), 8.76 (d, J=2.2 Hz, IH), 7.69 (s, IH), 4.25 (br s, 3H). Step 2: Preparation of furor2.3-b]pyridine-5-carboxylic acid To a suspension of furo[2,3-b]pyridine-2,5-dicarboxylic acid (0.36 g, 1.484 mmol) in 3 mL of quinoline, under Ar, was added Cu powder (180 mg, 2.82 mmol) and warmed to 210°C for 1.5 hours. The reaction was cooled to RT and diluted with 50 mL of methylene chloride and filtered through celite. The organic layer was extracted with sat'd NaCθ3 (2 x 40 mL), acidified to pH 3 with 3 N HCI, and filtered to afford 80 mg of a tan solid. The aqueous layer was extracted with ether/methanol (85/15) (3 x 50 mL) and washed with brine (1 x 10 mL), dried over MgSθ4, filtered and concentrated to afford an additional 35 mg of product. lH NMR (400 MHz, (CD3OD) δ 8.89 (s, IH), 8.67 (d, J=2.0 Hz, IH), 7.97 (d, J=2.5 Hz, IH), 7.01 (d, J=2.4 Hz, IH). Step 3: Preparation of methyl furor2.3-blpyridine-5-carboxylate To furo[2,3-b]pyridine-5-carboxylic acid (3.0 g, 18.40 mmol) dissolved in 40 mL of methanol was added 160 mL of chloroform and then trimethysilyldiazomethane (42 mL, 10% solution in hexanes) slowly. After 0.5 hours 4 drops glacial acetic acid was added and the reaction mixture was concentrated. This provided 3.20 g as an off white solid. lH NMR (400 MHz, CDCI3) δ 9.02 (d, J=2.0 Hz, IH), 8.60 (d, J=2.0 Hz, IH), 7.79 (d, J=2.5 Hz, IH), 6.87 (d, J=2.5 Hz, IH), 3.98 (s, 3H). Step 4: Preparation of 5-hvdroxymethyl furor2.3-b]pyridine A flame dried 500 mL round bottom flask was charged with methyl furo[2,3-b]pyridine-5-carboxylate (3.20 g, 18.08 mmol) dissolved in 90 mL of THF and cooled to 0°C. To this was added diisobutylaluminum hydride (46 mL, 46.1 mmol, 1 M solution in hexanes) over 10 minutes and the cooling bath removed. After 4 hours the reaction mixture was cooled to 0°C and slowly quenched with rochelle salts (100 mL). After an additional 18 hours the layers were separated and the aqueous layer was extracted with ethyl acetate (4 x 40 mL). The combined organic layers were washed with brine (1 x 20 mL), dried over MgSθ4, filtered and concentrated. The residue was purified via flash column chromatography (40 x 150 mm column, gradient ώlution CH2Cl2:CH2Cl2 sat'd with NH3:MeOH 60:39:1.0 (1000 mL), 60:38:2 (1000 mL), 60:37:3 (1000 mL), 60:36:4 (1000 mL). This provided 2.16 g of a white solid. lH NMR (400 MHz, CDCI3) δ 8.19 (d, J=2.0 Hz, IH), 7.92 (d, J=2.0 Hz, IH), 7.64 (d, J=2.5 Hz, IH), 6.69 (d, J=2.4 Hz, IH), 4.78 (d, J=3.8 Hz, 2H), 4.69 (br s, IH). Step 5: Preparation of 3 -chloromethyl furo[2,3-b]pyridine hydrochloride HCI To a solution of 5-hydroxymethyl furo[2,3-b]pyridine dissolved in 9 mL of methylene chloride cooled to 0°C was added thionyl chloride (4.23 mL, 57.99 mmol). The ice bath was removed and after 1 hour the reaction mixture was concentrated to afford 2.86 g of an off white solid. lH NMR (400 MHz, CDCI3) δ 8.40 (d, J=2.0 Hz, IH), 8.13 (d, J=2.2 Hz, IH), 7.80 (d, J=2.4 Hz, IH), 6.86 (d, J=2.4 Hz, IH), 4.74 (s, 2H). Step 6: Preparation of N-(2(R)-hydroxy-l(S)-indanyl)-2(R)- phenylmethyl-4(S)-hydroxy-5-(l-(4-(3-furo[2,3-b]- pyridylmethyl)-2(S)-N'-(t-butylcarboxamido)- piperazinyl))-pentaneamide To a solution of N-(2(R)-hydroxy-l(S)-indanyl)-2(R)- phenylmethyl-4(S)-hydroxy-5(-2(S)-N'-(t-butylcarboxamido)- piperazinyl))pentaneamide (6.50 g, 12.48 mmol) dissolved in 12 mL of dimethylformamide, under argon, was added 3-chloromethylfuro[2,3- b]pyridine hydrochloride (2.80 g, 13.72 mmol) and triethylamine (5.21 mL, 37.44 mmol). After 18 hours the reaction mixture was diluted with 400 mL of ethyl acetate and washed with sat'd NaHCθ3 (1 x 25 mL), water (5 x 20 mL), and brine (1 x 25 mL). The solution was dried over MgS04, filtered and concentrated to an oil. The residue was purified via flash column chromatography (60 x 150 mm column, gradient elution CH2d2:CH2Cl2 sat'd with NH3:MeOH 60:39:1.0 (1000 mL), 60:38:2 (1500 mL), 60:37:3 (1500 mL), 60:36:4 (1500 mL). Titrated the resulting foam in ethyl acetate and the desired product was filtered and dried overnight under high vacuum at 65°C to provide 5.30 g of white crystalline solid. Mixed fractions from the column chromatography could be combined and repurified to afford more product, mp 183.5-184.5°C. lH NMR (400 MHz, CDCI3) δ 8.25 (d, J=2.2 Hz, IH), 7.85 (d, J=2.0 Hz, IH), 7.75 (s, IH), 7.73 (d, J=2.4 Hz, IH), 7.32-7.10 (m, 9H), 6.75 (d, J=2.4 Hz, IH), 5.95 (d, J=8.6 Hz, IH), 5.27 (dd, J=8.5, and 4.8 Hz, IH), 4.27-4.26 (m, IH), 4.12 (br s, IH), 3.89-3.83 (m, IH), 3.51 (s, 2H), 3.29 (dd, J=17.5 and 4.0 Hz, IH), 3.16 (dd, J=3.66 and 3.48 Hz, IH), 3.15 (dd, J=6.6 and 5.1 Hz, IH), 2.94-2.50 (m, 11H), 2.36-2.34 (m, IH), 1.66 (s, IH), 1.62-1.47 (m, IH), 1.35 (s, 9H). Analysis calculated for C38H47N5O5 C, 69.81; H, 7.25; N, 10.71 Found: C, 69.46; H, 7.22; N, 10.69 EXAMPLE 3 Employing substantially the same procedure as described in Example 2, but treating the N-(2(R)-hydroxy-l(S)-indanyl)-2(R)- phenylmethyl-4(S)-hydroxy-5-(l-(2(S)-N'-(t-butylcarboxamido)- piperazinyl))-pentaneamide used therein (compound (i) below) with the alkylating agent (ii) indicated below in place of the alkylating agent used in Step 6 therein, the following products defined by formula (iii) were made: FT-X R η -X 25 30 EXAMPLE 4 Preparation of Amide 1 A solution of (-)-cis-l-aminoindan-2-ol (884 g, 5.93 mol) in 17.8 L of dry THF (KF = 55 mg/mL) (KF stands for Karl Fisher titration for water) and triethylamine (868 mL, 6.22 mol) in a 50 L round bottom flask equipped with a thermocouple probe, mechanical stirrer, and a nitrogen inlet adapter and bubbler, was cooled to 15°C. Then, 3-phenylpropionyl chloride (1000 g, 5.93 mol) was added over 75 minutes, while the internal temperature was kept between 14-24°C with an ice-water cooling batch. After addition, the mixture was aged at 18 to 20°C for 30 minutes and checked by HPLC analysis for the disappearance of (-)-cis-l-aminoindan-2-ol. Progress of the reaction is monitored by high performance liquid chromatography (HPLC) analysis: 25 cm Dupont C8-RX column, 60:40 acetonitrile/10 mM (KH2PO4/K2HPO4), 1.0 mL/min., injection volume = 20 mL, detection = 200 nm, sample preparation = 500 x dilution. Approximate retention times: retention time (min) identity 6.3 cis-aminoindanol The reaction was treated with pyridinium -toluene- sulfonate (241 g, 0.96 mol, 0.16 equiv.) and stirred for 10 minutes (the pH of the mixture after diluting 1 mL sample with an equal volume of water is between 4.3-4.6). Then, 2-methoxypropene (1.27 L, 13.24 mol, 2.2 equiv.) was added and reaction was heated to 38-40°C for 2 hours. The reaction mixture was cooled to 20°C and partitioned with ethyl acetate (12 L) and 5% aqueous NaHC03 (10 L). The mixture was agitated and the layers were separated. The ethyl acetate extract was washed with 5% aqueous NaHCθ3 (10 L) and water (4 L). The ethyl acetate extract was dried by atmospheric distillation and solvent switched to cyclohexane (total volume of ~30L). At the end of the distillation and concentration (20 volume % of ethyl acetate extraction volume), the hot cyclohexane solution was allowed to slowly cool to 25°C to crystallize the product. The resulting slurry was further cooled to 10°C and aged for 1 hour. The product was isolated by filtration and the wet cake was washed with cold (10°C) cyclohexane (2 x 800 mL). The washed cake was dried under vacuum (26" of Hg) at 40°C to afford 1.65 kg of acetonide 1 (86.4%, 98 area% by HPLC), lH NMR (300.13 MHz, CDCI3, major rotamer) δ 7.36-7.14 (m, 9 H), 5.03 (d, J=4.4, 1 H), 4.66 (m, 1 H) 3.15 (m, 2 H), 3.06 (br s, 2 H), 2.97 (m, 2 H), 1.62 (s, 3 H), 1.37 (s, 3 H); 13c NMR (75.5 MHz, CDCI3, major rotamer) δ c 168.8, 140.9, 140.8, 140.6, 128.6, 128.5, 128.4, 127.1, 126.3, 125.8, 124.1, 96.5, 78.6, 65.9, 38.4, 36.2, 31.9, 26.5, 24.1. Analysis calculated for C21 H23NO2 C, 78.47; H, 7.21; N, 4.36 Found: C, 78.65; H, 7.24; N, 4.40 EXAMPLE 5 Preparation of Epoxide 3 A solution of acetonide 1 (1000 g c , 3.11 mol) and 2(S)- glycidyl tosylate 2 (853 g, 3.74 mol, 1.2 equiv.) in 15.6 L of THF (KF = 22 mg/mL) in a 50 L 4-neck round bottom flask, equipped with a thermocouple, mechanical stirrer, addition funnel and nitrogen inlet adapter was degassed 3 times via vacuum-nitrogen purge and cooled to -56°C. Then, lithium hexamethyldisilazide (LiN[(CH3)3Si]2)(2.6 L, 1.38 M, 1.15 equiv.) was added over 2 hours, while keeping the internal temperature between -50 to -45 °C. The reaction mixture was stirred at -45 to -40°C for 1 hour and then allowed to warm to -25°C over 1 hour. The mixture is stirred between -25 to -22°C for 4 hours (or until the starting acetonide is 3.0 area %). Progress of the reaction is monitored by HPLC analysis: 25 cm x 4.6 nm Zorbax Silica column, 20% ethyl acetate in hexane, 2.0 mL/min, injection volume = 20 mL, detection = 254 nm, sample preparation = 100 x dilution. Approximate retention times: retention time (min) identity 5.5 amide 1 6.5 glycidyl tosylate 2 13.5 epoxide 3 The reaction mixture was quenched with DI water (6.7 L) at -15°Gand partitioned with ethyl acetate (10 L). The mixture was agitated and the layers were separated. The ethyl acetate extract was washed with a mixture of 1% aqueous NaHCθ3 (5 L) and saturated NaCl (0.5 L). The ethyl acetate extract (28.3 L) was concentrated by vacuum distillation (28" of Hg) and additional ethyl acetate was added to complete the solvent switch to ethyl acetate (final volume = 11.7 L). The ethyl acetate concentrate was further solvent switched to MeOH to crystallize the product and concentrated to a final volume of 3.2 L. The residual ethyl acetate solvent was removed by charging 10 L of methanol and collecting 10 L of distillate. The resulting slurry was stirred at 22°C for 1 hour, then cooled to 5°C and aged for 0.5 hours. The product was isolated by filtration and the wet cake was washed with cold methanol (2 x 250 mL). The washed cake was dried under vacuum (26" of Hg) at 25°C to afford 727 g of epoxide 3 (61.2%, 98.7 area % of the major epoxide by HPLC): 1 c NMR (300 MHz, CDCI3) δ 171.1, 140.6, 140.5, 139.6, 129.6, 128.8, 128.2, 127.2, 126.8, 125.6, 124.1 , 96.8, 79.2, 65.8, 50.0, 48.0, 44.8, 39.2, 37.4, 36.2, 26.6, 24.1. EXAMPLE 6 Preparation of penultimate 6 A slurry of the 2(S)-t-butylcarboxamide-4-/v r -Boc- piperazine 4 (1950 g, 6.83 mol, >99.5% ee) (ee = enantiomeric excess) and the epoxide 3 (2456 g, 97.5:2.5 mixture of 45/R epoxides, 6.51 mol) in isopropanol (2-propanol, 18.6 L) in a 72 L round bottom flask with four inlets, equipped with a mechanical stirrer, reflux condenser, steam bath, Teflon coated thermocouple and nitrogen inlet, was heated to reflux (internal temperature was 84-85°C). After 40 min, a homogeneous solution was obtained. The mixture was heated at reflux for 28 hours. The internal temperature during reflux was 84-85°C. Progress of the reaction was monitored by HPLC analysis: 25 cm Dupont C8-RX column, 60:40 acetonitrile/10 mM (KH2PO4/K2HPO4), 1.0 mL/min, detection = 220 nm, sample preparation = 2 μL, reaction mixture diluted to 1 mL in acetonitriie. Approximate retention times: retention time (min) identity 4.8 piperazine 4 8.9 epoxide 3 15.2 coupled product 5 After 28 hours, the remaining epoxide 3 and coupled product 5 (by HPLC analysis) were 1.5 area % and 91-93 area %, respectively. The mixture was cooled to 0 to 5°C and 20.9 L of 6 N HCI was added while keeping the temperature below 15°C. After the addition was complete, the mixture was warmed to 22°C. Evolution of gas is noted at this point (isobutylene). The mixture was aged at 20 to 22°C for 6 hours. Progress of the reaction was monitored by HPLC analysis: same conditions as above. Approximate retention times: retention time (min) identity 7.0 cis-aminoindanol 11.9 penultimate 6 15.1 coupled product 5 The mixture was cooled to 0°C and 7.5 L of 50% NaOH was slowly added to adjust the pH of the mixture to pH=l 1.6, while keeping the temperature less than 25°C during the addition. The mixture was partitioned with ethyl acetate (40 L) and water (3 L . The mixture was agitated and the layers were separated. The organic phase (60 L) was concentrated under reduced pressure (29" of Hg) and solvent s witched to DMF and concentrated to a final volume of 10.5 L (KF = 1.8 mg/mL). The HPLC assay yield of 6 in ethyl acetate was 86.5%. The penultimate Compound 6 in DMF was directly used in the next step without further purification. For isolated 6: 13c NMR (75.4 MHz, CDCI3) δ 175.2, 170.5, 140.8, 140.5, 139.9, 129.1, 128.5, 127.9, 126.8, 126.5, 125.2, 124.2, 73.0, 66.0, 64.8, 62.2, 57.5, 49.5, 47.9, 46.4, 45.3, 39.6, 39.3, 38.2, 28.9. EXAMPLE 7 Pyrazine-2-tert-butyl carboxamide 9 8 2-Pyrazinecarboxylic acid (8) 3.35 kg (27 mol) Oxalyl chloride 3.46 kg (27.2 mol) tert-Butylamine (KF = 460 μg/ml) 9.36 L (89 mol) EtOAc (KF = 56 μg/ml) 27 L DMF 120 mL 1 -Propanol 30 L The carboxylic acid 8 was suspended in 27 L of EtOAc and 120 mL of DMF in a 72 L 3-neck flask with mechanical stirring under N2 and the suspension was cooled to 2°C. The oxalyl chloride was added, maintaining the temperature between 5 and 8°C. The addition was completed in 5 hours. During the exothermic addition CO and C02 were evolved. The HCI that was formed remained largely in solution. A precipitate was present which is probably the HCL salt of the pyrazine acid chloride. Assay of the acid chloride formation was carried out by quenching an anhydrous sample of the reaction with t-butylamine. At completion <0.7% of acid 8 remained. The assay for completion of the acid chloride formation is important because incomplete reaction leads to formation of a bis-tert- butyl oxamide impurity. The reaction can be monitored by HPLC: 25 cm Dupont Zorbax RXC8 column with 1 mL/min flow and detection at 250 nm; linear gradient from 98% of 0.1 % aqueous H3PO4 and 2% CH3CN to 50% aqueous H3PO4 and 50% CH3CN at 30 min. Retention times: acid 8 = 10.7 min, amide 9 = 28.1 min. The reaction mixture was aged at 5°C for 1 hour. The resulting slurry was cooled to 0°C and the tert-butylamine was added at such a rate as to keep the internal temperature below 20°C. The addition required 6 hours, as the reaction was very exothermic. A small portion of the generated tert-butylammonium hydrochloride was swept out of the reaction as a fluffy white solid. The mixture was aged at 18°C for an additional 30 min. The precipitated ammonium salts were removed by filtration. The filter cake was washed with 12 L of EtOAc. The combined organic phases were washed with 6 L of a 3% NaHCθ3 and 2 x 2 L of saturated aq. NaCl. The organic phase was treated with 200 g of Darco G60 carbon and filtered through Solka Flok and the cake was washed with 4 L of EtOAc. Carbon treatment efficiently removed some purple color in the product. The EtOAc solution of 9 was concentrated at 10 mbar to 25% of the original volume. 30 L of 1-propanol were added, and the distillation was continued until a final volume of 20 L was reached. At this point, the EtOAc was below the limit of detection in the lH NMR (<1%). The internal temperature in this solvent change was <30°C. A 1-propanol/EtOAC solution of 3 was stable to reflux atatmospheric pressure for several days. Evaporation of an aliquot gave a tan solid m.p 87-88°C. 13c NMR (75 MHz, CDCI3, ppm) 161.8, 146.8,145.0, 143.8, 142.1, 51.0, 28.5. EXAMPLE 8 rac-2-tert-Butyl-carboxamide-piperazine 10 Materials Pyrazine-2-tert-butylcarboxamide 9 2.4 kg (13.4 mol) in 1- Propanol solution 12 L 20% Pd(OH)2/C 16 wt.% water 144 g. The pyrazine-2-tert-butylcarboxamide 9/1-propanol solution was placed into the 5 gal autoclave. The catalyst was added and the mixture was hydrogenated at 65°C at 40 psi (3 atm) of H2. After 24 hours the reaction had taken up the theoretical amount of hydrogen and GC indicated <1% of 9. The mixture was cooled, purged with N2 and the catalyst was removed by filtration through Solka Floe. The catalyst was washed with 2 L of warm 1- propanol. It was found that the use of warm 1-propanol during washing of the filter cake improved filtration and lowered the losses of product on the filter cake. The reaction was monitored by GC:30 m Megabore column, from 100°C to 160°C at 10°C/min, hold 5 min, then at 10°C/min to 250°C, retention times: 9 = 7.0 min, 10 = 9.4 min. The reaction could also be monitored by TLC with EtOAc/MeOH (50:50) as solvent and Ninhydrin as developing agent. Evaporation of an aliquot indicated that the yield over amidation and hydrogenation is 88% and that the concentration of 10 is 133g/L. Evaporation of an aliquot gave 10 as a white solid mp 150- 151°C; 13c NMR (75 MHz, D2θ, ppm) 173.5, 59.8, 52.0, 48.7, 45.0, 44.8, 28.7. EXAMPLE 9 (S)-2-tert-Butyl-carboxamide-piperazine bis (S)-Camphorsulfonic acid salt (S)-ll 10 11 Materials rac-2-tert-Butyl-carboxamide-piperazine 10 4.10 kg (22.12 mol) in 1-Propanol Solution in 25.5 Kg solvent (S)-(+)-10-Camphorsulfonic acid 10.0 Kg (43.2 mol) 1-Propanol 12 L Acetonitriie 39 L Water 2.4 L The solution of amine 10 in 1-propanol was charged to a 100 L flask with an attached batch concentrator. The solution was concentrated at 10 mbar and a temperature <25°C to a volume of ca 12 L. At this point the product had precipitated from the solution, but went back into a solution when the mixture was heated to 50°C. Analysis of a homogeneous aliquot indicated that the concentration of 10 was 341 g/L. The concentration was determined by HPLC: 25 cm Dupont Zorbax RXC8 column with 1.5 mL/min flow and detection at 210 nm, isocratic (98/2) CH3CN/0.1% aqueous H3PO4. Retention time of 10 : 2.5 min. Acetonitriie (39 L) and water (2.4 L) were added to give a clear, slightly brown solution. Determination of the water content by KF titration and CH3CN/l-propanol ratio by lH NMR integration showed that the CH3CN/l-propanol/H2θ ratio was 26/8/1.6. The concentration in the solution was 72.2 g/L. The (S)-lO-camphorsulfonic acid was charged over 30 min in 4 portions at 20°C. The temperature rose to 40°C after the CSA was added. After a few minutes a thick white precipitate formed. The white slurry was heated to 76°C to dissolve all the solids, the slightly brown solution was then allowed to cool to 21 °C over 8 hours. The product precipitated at 62°C. The product was filtered without aging at 21 °C, and the filter cake was washed with 5 L of the CH3CN/l-propanol/H2θ 26/8/1.6 solvent mixture. It was dried at 35°C in the vacuum oven with N2 bleed to give 5.6 Kg (39%) of 11 as a white crystalline solid m.p 288-290°C (with decomp.). [ ]D 25 = 18.9° (c = 0.37, H2O). 13c NMR (75 MHz, D2O, ppm) 222.0, 164.0, 59.3, 54.9, 53.3, 49.0, 48.1, 43.6, 43.5, 43.1, 40.6, 40.4, 28.5, 27.2, 25.4, 19.9, 19.8. The ee of the material was 95% according to the following chiral HPLC assay: an aliquot of 11 (33 mg) was suspended in 4 mL of EtOH and 1 mL of Et3N. Boc2θ (11 mg) was added and the reaction mixture was allowed to age for 1 hour. The solvent was completely removed in vacuo, and the residue was dissolved in ca. 1 mL of EtOAc and filtered through a Pasteur pipet with Siθ2, using EtOAc as eluent. The evaporated product fractions were redissolved in hexanes at ca. 1 mg/mL. The enantiomers were separated on a Daicel Chiracell AS column with a hexane/IPA (97:3) solvent system at a flow rate of 1 mL/min and detection at 228 nm. Retention times: S antipode = 7.4 min, R = 9.7 min. EXAMPLE 10 (S)-2-tert-Butylcarboxamide-4-tert-butoxycarbonyl-piperazine 4 from salt 11 11 Materials (S)-2-tert-Butyl-carboxamide-piperazine Bis (S) - (+) - CSA salt 11, 95% ee 5.54 Kg (8.53 mol) Di-tert-butyl dicarbonate 1.86 Kg (8.53 mol) Et3N 5.95L (42.6 mol) EtOH Punctilious 200 proof 55 L EtOAc 2 L To the (S)-CSA salt 11 in a 100 L 3-neck flask with an addition funnel under N2 was added EtOH, followed by triethylamine at 25°C. The solid dissolved readily on the addition of the Et3N. The Boc2θ was dissolved in EtOAc and charged to the addition funnel. The solution of Boc2θ in EtOAc was added at such a rate as to keep the temperature below 25°C. The addition took 3 hours. The reaction mixture was aged for 1 hour after completion of the addition of the Boc2θ solution. The reaction can be monitored by HPLC: 25 cm Dupont Zorbax RXC8 column with 1 mL/min flow and detection at 228 nm, isocratic (50/50) CH3CN/0.1 M KH2PO4 adjusted to pH=6.8 with NaOH. Retention time of 4 = 7.2 min. The chiral assay was carried out using the same system as in the previous step. The reaction could also be monitored by TLC with a 100% EtOAc as the solvent. (Rf=0.7) The solution was then concentrated to ca. 10 L at an internal temperature of <20°C in a batch-type concentrator under 10 mbar vacuum. The solvent switch was completed by slowly bleeding in 20 L of EtOAc and reconcentrating to ca 10 L. The reaction mixture was washed into an extractor with 60 L of EtOAc. The organic phase was washed with 16 L of 5% aqueous Na2Cθ3 solution, 2 x 10 L Di water and 2 x 6 L of saturated aqueous sodium chloride. The combined aqueous washes were back extracted with 20 L of EtOAc and the organic phase was washed with 2 x 3 L water and 2 x 4 L of saturated aqueous sodium chloride. The combined EtOAc extracts were concentrated under 10 mbar vacuum with an internal temperature of <20°C in a 100 L batch-type concentrator to ca. 8 L. The solvent switch to cyclohexane was achieved by slowly bleeding in ca. 20 L of cyclohexane, and reconcentrating to ca. 8 L. To the slurry was added 5 L of cyclohexane and 280 mL of EtOAc and the mixture was heated to reflux, when everything went into solution. The solution was cooled and seed (10 g) was added at 58°C. The slurry was cooled to 22°C in 4 hours and the product was isolated by filtration after a 1 hour age at 22°C. The filter cake was washed with 1.8 L of cyclohexane and dried in the vacuum oven at 35°C under N2 bleed to give 1.87 Kg (77%, >99.9 area % by HPLC, R-isomer below level of detection) of 4 as a slightly tan powder. [ ]D 25 = 22.0° (c = 0.20, MeOH), m.p 107°C; 13c NMR (75 MHz, CDC13, ppm) 170.1, 154.5, 79.8, 58.7, 50.6, 46.6, 43.6, 43.4, 28.6, 28.3. While the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that the practice of the invention emcompasses all of the usual variations, adaptations, or modifications, as come within the scope of the following claims and its equivalents.

Claims: WHAT IS CLAIMED IS: 1. A compound of the formula: or a pharmaceutically acceptable salt thereof, wherein: -O is a stable 8- to 10-membered bicyclic heterocycle, any ring of which may be saturated or unsaturated, and said heterocycle consists of carbon atoms and 1-3 heteroatoms selected from the group consisting of N, S or O, said heterocycle unsubstituted or substituted with OH, halo, lower Cl-4 alkyl, oxo; with the proviso that ^- is neither 2. A compound of Claim 1, wherein: ■ o is a stable 8- to 10-membered bicyclic heterocycle, any ring of which may be saturated or unsaturated, and said heterocycle consists of carbon atoms and 2 heteroatoms selected from the group consisting of N or O, wherein the heteroatoms are in different rings, or a pharmaceutically acceptable salt thereof. A compound of Claim 1 , wherein: and X is O or S, or a pharmaceutically acceptable salt thereof. A compound of Claim 1 , wherein: -O is restricted to or a pharmaceutically acceptable salt thereof A compound of Claim 1, which is named N-(2(R)-hydroxy- 1 (S)-indanyl)-2(R)-phenylmethyl-4(S)- hydroxy-5-(l-(4-(3-furo[2,3-b]pyridylmethyl)-2(S)-N'-(t-butylcarbox- amido)-piperazinyl))pentaneamide, or a pharmaceutically acceptable salt thereof. 6. A pharmaceutical composition comprising a compound of any of Claims 1-5, and a pharmaceutically acceptable carrier. 7. The pharmaceutical composition of any of Claims 1-5, for use in the treatment of AIDS, in the prevention of infection by HTV, in the treatment of infection of HTV, or in the inhibition of HIV protease. 8. A method of treating AIDS, comprising administering to a mammal in need of such treatment an effective amount of a compound of any of Claims 1-5. 9. A method of preventing infection by HIV, comprising administering to a mammal in need of such treatment an effective amount of a compound of any of Claims 1-5. 10. A method of treating infection by HTV, comprising administering to a mammal in need of such treatment an effective amount of a compound of any of Claims 1-5. 11. A method of inhibiting HIV protease, comprising administering to a mammal in need of such treatment an effective amount of a compound of any of Claims 1-5. 12. A combination of compounds which is the compound of Claim 5, and a non-nucleoside analog inhibitor of HIV reverse transcriptase selected from Compound B and Compound C and nevirapine, and, optionally, any of AZT or ddl or ddC. 13. A combination of compounds which is the compound of Claim 5 and any of AZT or ddl or ddC.

Inventors: DORSEY BRUCE D, HUFF JOEL R, VACCA JOSEPH P, DORSEY, BRUCE, D., HUFF, JOEL, R., VACCA, JOSEPH, P.
Applicants: MERCK & CO INC, MERCK & CO., INC.
Application year: 1994
Main classification: C07D473
IPC codes: A61K31, A61P31, A61P37, C07D241, C07D307, C07D491, C07D495